Production of active pediocin PA-1 in Escherichia coli using a thioredoxin gene fusion expression approach: cloning, expression, purification, and characterization.

摘要

Antimicrobial peptides possess cationic and amphipathic properties that allow for interactions with the membrane of living cells. Bacteriocins from lactic acid bacteria, in particular, are currently being studied for their potential use as food preservatives and in health care. However, bacteriocin exploitation is often limited owing to low production yields. Gene cloning and heterologous protein or peptide production may be one way to overexpress bacteriocins. In this study, production of recombinant active pediocin PA-1 (PedA) was accomplished in Escherichia coli using a thioredoxin (trx) gene fusion (trx-pedA) expression approach. The recombinant protein Trx-PedA did not show any biological activity; however, upon cleavage by an enterokinase, biologically active pediocin PA-1 was obtained. Recombinant pediocin PA-1 characteristics (mol. wt., biological activity, physicochemical properties) were found to be very similar to those of native pediocin PA-1. In addition, a 4-5-fold increase in production yield was obtained, compared with PA-1 produced naturally by Pediococcus acidilactici PAC 1.0. It is concluded that the developed method, although not optimized, offers great potential for the production of pediocin PA-1 for high-value applications.